Tools towards the rapid derivation of glial cells from human pluripotent cells

从人类多能细胞中快速衍生出神经胶质细胞的工具

基本信息

批准号：
8665502
负责人：
LORENZ P. STUDER
金额：
$ 21.88万
依托单位：
SLOAN-KETTERING INST CAN RESEARCH
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-07-01 至 2015-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8665502
关键词：
Acceleration Affect Amyotrophic Lateral Sclerosis Animals Astrocytes Brain Cell Line Cells Cellular biology Data Derivation procedure Development Disease Disease model ES Cell Line Embryonic Development Engraftment Epidermal Growth Factor Receptor Epigenetic Process Fibroblasts Gene Expression Profile Gene Targeting Generations Glial Differentiation Glial Fibrillary Acidic Protein Goals Human In Vitro Light Mediating MicroRNAs Modeling Neuraxis Neurodegenerative Disorders Neurodevelopmental Disorder Neuroglia Neuronal Differentiation Neurons Oligodendroglia Pathogenesis Patients Phase Phenotype Population Pregnancy Process Production Property Proteins Protocols documentation Recording of previous events Regenerative Medicine Reporter Reporting Research Design Rett Syndrome Rodent Running Second Pregnancy Trimester Somatic Cell Staging Surrogate Markers Technology Testing Third Pregnancy Trimester Time Transcription Coactivator aquaporin 4 base cell type developmental neurobiology embryonic stem cell fetal genome-wide human disease human embryonic stem cell human embryonic stem cell line in vivo induced pluripotent stem cell insight interest multidisciplinary nerve stem cell nervous system development novel novel strategies nuclease pluripotency programs public health relevance receptor expression relating to nervous system screening self-renewal small hairpin RNA small molecule stem cell technology tool transcription factor

项目摘要

DESCRIPTION (provided by applicant): Neural stem cells (NSCs) are defined by their ability to self-renew and to differentiate into neurons, astrocytes and oligodendrocytes. However, during CNS development, the generation of neurons and glia is temporally regulated. In early embryonic development, NSCs are restricted and predominantly differentiate into neurons. Transitioning from early to late gestation, NSCs become competent towards glial differentiation. This suggests the presence of an epigenetic switch triggering the onset of glial production. Timing of the switch can be recapitulated in vitro; using primary or embryonic stem cell (ESC) derived NSCs. In human ESCs the switch towards efficient glial cell production occurs at 2-3 months after differentiation, a protracted time frame that represents a major practical hurdle for the application of glial cells in disease modeling and regenerative medicine. Here we propose to identify the mechanisms involved in the switch from neurogenic to gliogenic NSCs. Towards this goal we will establish a glial specific reporter human ESC lines targeting the glial fibrillary aciic protein (GFAP) and the aquaporin 4 (AQP4) to identify astrocytes and glial competent NSCs. Our preliminary results suggest that the several factors, including transcription factors as well a microRNA regulators, are differentially regulated in early versus late NSCs. Initially we will functionally test our candidate proteins for the ability to activate the gliogenic program in early NSCs. The reporter cell lines will serve as readout for a large-scale shRNA screen, aimed at identifying novel candidates that mediate the epigenetic switch in NSCs. Additionally, we found that the epidermal growth factor receptor (EGFR) is expressed in NSCs that correlate with glial competency. We will utilize EGFR to distinguish gliogenic NSCs (EGFR+ GFAP+) from glial cells (EGRF- GFAP+). Overall, the proposed studies are designed to yield novel insights into CNS fate choice and further our understanding of glial cell biology, ultimately identifying factors that may accelerate their differentiation from human pluripotent cells.

描述（由申请人提供）：神经干细胞（NSC）的定义是其自我更新和分化为神经元、星形胶质细胞和少突胶质细胞的能力。然而，在中枢神经系统发育过程中，神经元和神经胶质细胞的生成受到暂时调节。在早期胚胎发育中，神经干细胞受到限制并主要分化为神经元。从妊娠早期过渡到妊娠晚期，神经干细胞变得有能力进行神经胶质分化。这表明表观遗传开关的存在触发了神经胶质细胞的产生。开关的时间可以在体外重现；使用原代或胚胎干细胞 (ESC) 衍生的 NSC。在人类ESC中，分化后2-3个月发生向有效神经胶质细胞产生的转变，这是一个漫长的时间范围，代表了神经胶质细胞在疾病建模和再生医学中应用的主要实际障碍。在这里，我们建议确定从神经源性 NSC 转变为胶质源性 NSC 所涉及的机制。为了实现这一目标，我们将建立一个针对神经胶质原纤维酸性蛋白（GFAP）和水通道蛋白4（AQP4）的神经胶质特异性报告人类ESC系，以识别星形胶质细胞和神经胶质细胞。我们的初步结果表明，包括转录因子和 microRNA 调节因子在内的多种因子在早期和晚期 NSC 中受到差异性调节。最初，我们将对候选蛋白进行功能测试，以确定其在早期激活神经胶质生成程序的能力。国家干细胞。报告细胞系将作为大规模 shRNA 筛选的读数，旨在识别介导 NSC 表观遗传开关的新候选者。此外，我们发现表皮生长因子受体（EGFR）在与神经胶质细胞能力相关的神经干细胞中表达。我们将利用 EGFR 来区分胶质细胞 (EGFR+ GFAP+) 和神经胶质细胞 (EGRF- GFAP+)。总体而言，拟议的研究旨在对中枢神经系统命运选择产生新的见解，并进一步加深我们对神经胶质细胞生物学的理解，最终确定因素这可能会加速它们与人类多能细胞的分化。